Iba1 (AIF1) Mouse Monoclonal Antibody [Clone ID: 1022-5]

Specifications

Product Data
Clone Name	1022-5
Applications	IHC
Recommend Dilution	Immunohistochemistry on Paraffin Sctions: 10-20 μg/ml (1/20-1/0). Mcrowave pretreatment for antigen retrieval recommended. Suggested Positive Control: Human spleen. Does not react on routinely processed Frozen Sections.
Reactivity	Human, Porcine, Rat
Host	Mouse
Clonality	Monoclonal
Immunogen	Purified Human recombinant AIF-1. Epitope not further characterized.
Specificity	The antibody reacts with AIF-1.
Formulation	PBS, pH 7.2 containing 5 mg/ml BSA as stabilizer and 0.05% (v/v) Kathon as preservative State: Aff - Purified State: Lyophilized purified IgG fraction
Reconstitution Method	Restore by adding 0.5 ml distilled water.
Concentration	0.4 mg/ml (after reconstitution)
Purification	Affinity Chromatography
Conjugation	Unconjugated
Storage Condition	Store lyophilized at 2-8°C for 6 months or at -20°C long term. After reconstitution store the antibody undiluted at 2-8°C for one month  or (in aliquots) at -20°C long term. Avoid repeated freezing and thawing.
Gene Name	allograft inflammatory factor 1
Database Link	Entrez Gene 199 Human UniProt ID P55008
Background	Allograft Inflammatory Factor-1 is a Ca2+-binding peptide produced by activated macrophages and microglial cells. It has been suggested that AIF-1 expression is associated with chronic inflammatory processes. AIF-1, like the Ca2+-binding peptides MRP8 and MRP14, is expressed by activated macrophages and might participate in a variety of pathogenic processes in the mammalian brain and in chronic transplant rejection. It has been shown to be expressed early and persistently in chronically rejecting cardiac allografts but not in cardiac syngrafts and host hearts. The recently described apoptosis-inducing factor AIF is not related to AIF-1.
Synonyms	AIF-1, Protein G1, IBA1, Allograft inflammatory factor 1
Note	Protocol: Immunohistochemitry With Unlabeled Monoclonal Antibodies On Paraffin-Embedded Tissue Sections 1. Deparaffinize slides in chloroform for 30min and rehydrate to distilled water 2. Antigen retrieval: microwave treatment (700W) for 15min in 300ml 0.01M citrate buffer pH 6.0 3. Incubate slides for 15min in 200ml methanol/4.8ml 30%H2O2 4. Rinse slides 2x in distilled water and for 5min in TBS, pH 7.5 5. Block with normal swine serum diluted 1:10 in Tris buffered saline pH 7.5, containing 0.1% BSA. Cover sections with NSS and incubate for 15min in a humid chamber at RT  6. Remove serum from sections (do not rinse) 7. Incubate with primary antibody diluted in TBS/0.1%BSA, pH 7.5 for 60min in a humid chamber at RT  8. Rinse sections in TBS, pH 7.5 for 5min, 2x 9. Incubate with secondary antibody (Rabbit anti Mouse IgG+IgM-Biotin) 1:400 diluted in TBS/0.1%BSA, pH 7.5 for 30min in a moist chamber, RT. Preincubate now StreptABComplex /HRPO in TBS/0.1%BSA, pH 7.5! 10. Rinse sections in TBS, pH 7.5 for 5min, 2x 11. Incubate with StreptABComplex /HRPO for 30min in a humid chamber at RT   12. Rinse sections in TBS, pH 7.5 for 5min, 2x 13. DAB-staining Prepare 3,3' - diaminobenzidintetrahydrochloride (DAB) according the manufacturers instructions (e.g. DAB tablets) and incubate slides for 1-15 min. Check color reaction to avoid overstaining. 14. Rinse sections in TBS, pH 7.5 for 5min, 2x 15. Counterstain in haematoxylin for 1min and rinse in cold tap water for 10min 16. Mount slides with coverslips
Reference Data

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